electroporation

[ZEGSUS]

I want to make electroporation but I donit know from which microbе to extract DNA and to which to insert DNA.
Which combination is the most easy?

[JackBean]

E.coli -> E.coli

but I don't see, why would you do that just to do that.

[ZEGSUS]

And is it possible bacillus bulgaricus -->e.coli?

[JackBean]

sure, you can do salmon's DNA into acetobacter. It's just questionable, whether will the DNA stay or will be degraded or what you want to do with it (but I guess nothing since you don't care about the organisms ).

[ZEGSUS]

Well as you know I want to extract DNA from bulgaricus and to insert it in E.coli.The DNA will be in ethanol in the fridge.

[JackBean]

yeah, I understood that But what DNA will it be? Plasmid or gDNA?

But I guess you want to clone only small part and insert into some plasmid and express?

[ZEGSUS]

I will use and the both type because I haven't equipment to separate them.No , I will not clone them , I will try directly to insert the DNA into the E.coli.

[JackBean]

you have tuo purify the DNA first and that's were you differentiate between gDNA and plasmid. However, plasmid may be incompatible because of other replication origin. Inserting gDNA doesn't make sense since bacteria cannot have two chromosomes and thus would one repel probably (even keeping the plasmid without antibiotic selection will be hard).

[ZEGSUS]

And how to insert gDNA into the chromosome?

[JackBean]

you could use BAC (bacterial artificial chromosome), which has high capacity, but still I'm not sure, whether will it fit whole genome of your bacteria.
You want to create gDNA library?

[ZEGSUS]

And how to use BAC?

[JackBean]

I have never worked with BAC, but I guess cut with RE, insert your DNA and transform. Although I don't know whether electroporation will work for such a big piece.